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Sucrose Fluorometric Assay Kit
SKU: E-BC-F042-96
Sucrose Fluorometric Assay Kit
| SKU # | E-BC-F042 |
| Detection Instrument | Fluorescence microplate reader (Ex/Em=535 nm/590 nm) |
| Detection Method | Fluorescence method |
Product Details
Properties
| Sample Type | Plant tissue |
| Sensitivity | 0.15 μmol/L |
| Detection Range | 0.15-15 μmol/L |
| Detection Method | Fluorescence method |
| Assay Time | 70 min |
| Precision | inter-assay CV: 6.5% | intra-assay CV: 2.3% |
| Storage | This product can be stored at -20°C for 12 months with shading light. |
| Valid period | 12 months |
Dilution Of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
|---|---|
| 10% Corn tissue homogenate | 1500 |
| 10% Potato tissue homogenate | 300-500 |
| 10% Tomato tissue homogenate | 200-300 |
| 10% Macrophanerophytes leaf tissue homogenate | 200-400 |
| 10% Carrot tissue homogenate | 1500 |
| 10% Onion tissue homogenate | 500-1000 |
| 10% Green pepper tissue homogenate | 500-1000 |
| 10% Bush leaves tissue homogenate | 20-50 |
The diluent is extraction working solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Sucrose can be hydrolyzed by sucrase to produce glucose under acidic conditions, which is catalyzed by glucose oxidase to produce hydrogen peroxide. In the presence of HRP (horse radish peroxidase), hydrogen peroxide reacts with the fluorescent probe to form red fluorescent substance. The sucrose content can be calculated by measuring the fluorescence intensity at the excitation wavelength of 535 nm and the emission wavelength of 590 nm.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Extraction Solution | 45 mL × 1 vial | -20℃, 12 months |
| Reagent 2 | Enzyme Reagent 1 | Powder × 1 vial | -20℃, 12 months shading light |
| Reagent 3 | Buffer Solution | 10 mL × 1 vial | -20℃, 12 months |
| Reagent 4 | Enzyme Reagent 2 | Powder × 1 vial | -20℃, 12 months shading light |
| Reagent 5 | Probe | 0.25 mL × 1 vial | -20℃, 12 months shading light |
| Reagent 6 | Standard | Powder × 1 vial | -20℃, 12 months |
| Black Microplate | 96 wells | No requirement | |
| Plate Sealer | 2 pieces |
Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.
Technical Data:
Parameter:
Intra-assay Precision
Three tomato tissue samples were assayed in replicates of 20 to determine precision within an assay. (CV = Coefficient of Variation)
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μmol/L) | 1.00 | 7.00 | 13.00 |
| %CV | 2.5 | 2.2 | 2.2 |
Inter-assay Precision
Three tomato tissue samples were assayed 20 times in duplicate by three operators to determine precision between assays
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μmol/L) | 1.00 | 7.00 | 13.00 |
| %CV | 6.4 | 6.8 | 6.3 |
Recovery
Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 96%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (μmol/L) | 1.5 | 7.5 | 10 |
| Observed Conc. (μmol/L) | 1.5 | 7.1 | 9.5 |
| Recovery rate (%) | 98 | 95 | 9.5 |
Sensitivity
The analytical sensitivity of the assay is 0.15 μmol/L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.
Standard Curve:
As the fluorescence value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:
| Concentration (μmol/L) | 0 | 1 | 2 | 5 | 8 | 10 | 12 | 15 |
| Fluorescence value | 3020 | 4234 | 4953 | 8142 | 11568 | 13374 | 15512 | 18765 |
| 3130 | 4268 | 5212 | 8319 | 11609 | 13484 | 15714 | 18776 | |
| Average fluorescence value | 3075 | 4251 | 5083 | 8231 | 11589 | 13429 | 15613 | 18771 |
| Absoluted fluorescence value | 0 | 1176 | 2008 | 5156 | 8514 | 10354 | 12538 | 15696 |